Requirements for activation of human peripheral blood T cells by mouse monoclonal antibodies to CD3

Umetsu, Dale T.; Katzen, D; Chatila, Talal A.; Miller, Richard W.; Jabara, Haifa H.; Maher, Maureen; Oettgen, Hans C.; Terhorst, Cox; Geha, Raif S.

doi:10.1016/0090-1229(87)90156-5

“…Soluble factors, such as cytokines, and interaction with cell surface receptors have both been implicated as co-stimulators in TClUCD3-mediated activation [2]. Of the latter group, the adhesion molecules have recently attracted much attention [3].…”

Section: Engagement Of the T Cell Antigen Receptor (Tcr)/cd3mentioning

confidence: 99%

The role of lymphocyte subsets and adhesion molecules in T cell‐dependent cytotoxicity mediated by CD3 and CD28 bispecific monoclonal antibodies

Renner

¹

,

Jung

²

,

Şahin

³

et al. 1995

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The cure of human Hodgkin's tumors heterotransplanted into SCID mice can be achieved by two bispecific monoclonal antibodies (Bi-mAb) directed against the tumor-associated CD30 antigen and CD3 and CD28, respectively, and normal peripheral human blood T cells. We investigated the role of lymphocyte subsets and adhesion molecules in this Bi-mAb-mediated cytolysis. CD4+ lymphocytes were the most rapidly expanding subpopulation, but Bi-mAb-directed cytotoxicity was mediated preferentially by CD8+ lymphocytes and effector cells belonging to the CD45RO+ "memory" pool. Blocking of the LFA-1/ICAM-1 or CD2/LFA-3 adhesion pathways by mAb decreased Bi-mAb-mediated cytotoxicity. This was not due to inhibition of aggregate formation between Bi-mAb-coated T lymphocytes and target cells. Cross-linking of LFA-1 or CD2 molecules on lymphocytes prestimulated with Bi-mAb bound to CD3 and CD28 antigen lead to a more pronounced and prolonged rise in the intracellular concentration of free Ca2+. Additional CD2 cross-linking resulted in the tyrosine phosphorylation of distinct proteins. These findings indicate that adhesion molecules play a critical role and function as co-stimulatory signals rather than as cellular contact mediators in CD3 and CD28 Bi-mAb-stimulated T lymphocytes.

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“…Normal human fibroblasts were grown from skin biopsies, propagated, and harvested as described (20). Interferon y (IFN-y) treatment of fibroblasts was performed as described (20) L-Cell Transfectants.…”

Section: Introductionmentioning

confidence: 99%

Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules.

Scholl

¹

,

Diez

²

,

Mourad

³

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

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Toxic shock syndrome toxin 1 (TSST-1) is a 22-kDa exotoxin produced by strains of Staphylococcus aureus and implicated in the pathogenesis of toxic shock syndrome. In common with other staphylococcal exotoxins, TSST-1 has diverse immunological effects. These include the induction of interleukin 2 receptor expression, interleukin 2 synthesis, proliferation of human T lymphocytes, and stimulation of interleukin 1 synthesis by human monocytes. Toxic shock syndrome is a severe and often fatal disorder consistently associated with Staphylococcus aureus infection and characterized by multiple organ dysfunction (1-3). The exotoxin toxic shock syndrome toxin 1 (TSST-1) is a 22-kDa protein (4) produced by most strains of S. aureus isolated from patients with toxic shock syndrome (5, 6) and capable of inducing the disease in animal models (7). TSST-1 induces interleukin 1 synthesis by human monocytes (8, 9), accessory-cell-dependent proliferation of human T lymphocytes (10, 11), and the suppression of pokeweed mitogeninduced immunoglobulin synthesis by human lymphocytes (12). All three activities appear to reside in a 14-kDa internal toxin fragment and could not be dissected with multiple monoclonal antibodies (mAbs) (13,14). The identification of a cellular receptor for TSST-1 is important for the understanding of the mechanism of action of this toxin. TSST-1 has been reported to bind to cultured epithelial cells (15) and to T cells but not to monocytes or B cells (16). To the best of our knowledge, the nature of the binding site has not been described. In the present study we demonstrate that TSST-1 binds to human major histocompatibility complex (MHC) class II molecules.MATERIALS AND METHODS Cell Preparations. Peripheral blood mononuclear cells (PBMCs) were isolated by standard methods (11). Highly purified populations of T lymphocytes and monocytes were prepared as described (11,17). The preparation of B cells (18) from surgical specimens of human tonsil and of thymocytes (19) obtained from surgical specimens removed during open heart surgery were as described.Normal human fibroblasts were grown from skin biopsies, propagated, and harvested as described (20). Interferon y (IFN-y) treatment of fibroblasts was performed as described (20) L-Cell Transfectants. The L-cell transfectants expressing HLA class II molecules used in this study were gifts of R. Karr (University of Iowa, IA) and were prepared by transfection of the respective full-length cDNA clones (DRa, DR4f3I, DPw4a, DPw4p, DQ7a, and DQw3,8) in the expression vector pcD together with the pSV2-neo plasmid into DAP.3 murine L-cell fibroblasts (21). Cells were grown in tissue culture plates in complete RPMI 1640 medium containing G418 (250 jug/ml; GIBCO, Grand Island, NY) and were harvested by incubation for 15-20 min in isotonic phosphate-buffered saline (pH 7.4; PBS) containing 0.53 mM EDTA.Toxins. TSST-1 was prepared as described (22) and iodinated using chloramine-T. The mean specific activity of the labeled toxin was 950 Ci/mmol (range, 230-1700 Ci/m...

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“…Peripheral blood mononuclear cells (PBMCs) were isolated by standard methods (11). Highly purified populations of T lymphocytes and monocytes were prepared as described (11,17). The preparation of B cells (18) from surgical specimens of human tonsil and of thymocytes (19) obtained from surgical specimens removed during open heart surgery were as described.…”

Section: Introductionmentioning

confidence: 99%

“…Highly purified populations of T lymphocytes and monocytes were prepared as described (11,17). The preparation of B cells (18) from surgical specimens of human tonsil and of thymocytes (19) obtained from surgical specimens removed during open heart surgery were as described.Normal human fibroblasts were grown from skin biopsies, propagated, and harvested as described (20). Interferon y (IFN-y) treatment of fibroblasts was performed as described (20) L-Cell Transfectants.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules

1989

Proc. Natl. Acad. Sci. U.S.A.

2

0

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Toxic shock syndrome toxin 1 (TSST-1) is a 22-kDa exotoxin produced by strains of Staphylococcus aureus and implicated in the pathogenesis of toxic shock syndrome. In common with other staphylococcal exotoxins, TSST-1 has diverse immunological effects. These include the induction of interleukin 2 receptor expression, interleukin 2 synthesis, proliferation of human T lymphocytes, and stimulation of interleukin 1 synthesis by human monocytes. Toxic shock syndrome is a severe and often fatal disorder consistently associated with Staphylococcus aureus infection and characterized by multiple organ dysfunction (1-3). The exotoxin toxic shock syndrome toxin 1 (TSST-1) is a 22-kDa protein (4) produced by most strains of S. aureus isolated from patients with toxic shock syndrome (5, 6) and capable of inducing the disease in animal models (7). TSST-1 induces interleukin 1 synthesis by human monocytes (8, 9), accessory-cell-dependent proliferation of human T lymphocytes (10, 11), and the suppression of pokeweed mitogeninduced immunoglobulin synthesis by human lymphocytes (12). All three activities appear to reside in a 14-kDa internal toxin fragment and could not be dissected with multiple monoclonal antibodies (mAbs) (13,14). The identification of a cellular receptor for TSST-1 is important for the understanding of the mechanism of action of this toxin. TSST-1 has been reported to bind to cultured epithelial cells (15) and to T cells but not to monocytes or B cells (16). To the best of our knowledge, the nature of the binding site has not been described. In the present study we demonstrate that TSST-1 binds to human major histocompatibility complex (MHC) class II molecules.MATERIALS AND METHODS Cell Preparations. Peripheral blood mononuclear cells (PBMCs) were isolated by standard methods (11). Highly purified populations of T lymphocytes and monocytes were prepared as described (11,17). The preparation of B cells (18) from surgical specimens of human tonsil and of thymocytes (19) obtained from surgical specimens removed during open heart surgery were as described.Normal human fibroblasts were grown from skin biopsies, propagated, and harvested as described (20). Interferon y (IFN-y) treatment of fibroblasts was performed as described (20) L-Cell Transfectants. The L-cell transfectants expressing HLA class II molecules used in this study were gifts of R. Karr (University of Iowa, IA) and were prepared by transfection of the respective full-length cDNA clones (DRa, DR4f3I, DPw4a, DPw4p, DQ7a, and DQw3,8) in the expression vector pcD together with the pSV2-neo plasmid into DAP.3 murine L-cell fibroblasts (21). Cells were grown in tissue culture plates in complete RPMI 1640 medium containing G418 (250 jug/ml; GIBCO, Grand Island, NY) and were harvested by incubation for 15-20 min in isotonic phosphate-buffered saline (pH 7.4; PBS) containing 0.53 mM EDTA.Toxins. TSST-1 was prepared as described (22) and iodinated using chloramine-T. The mean specific activity of the labeled toxin was 950 Ci/mmol (range, 230-1700 Ci/m...

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Requirements for activation of human peripheral blood T cells by mouse monoclonal antibodies to CD3

Cited by 32 publications

References 34 publications

The role of lymphocyte subsets and adhesion molecules in T cell‐dependent cytotoxicity mediated by CD3 and CD28 bispecific monoclonal antibodies

The role of lymphocyte subsets and adhesion molecules in T cell‐dependent cytotoxicity mediated by CD3 and CD28 bispecific monoclonal antibodies

Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules.

Toxic shock syndrome toxin 1 binds to major histocompatibility complex class II molecules

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